ULTRA-SENSITIVE DETECTION METHODOLOGIES FOR SUBSTANCE P

这项研究提案的最终目标是开发一种测量高离子生物液体中P物质(SP)及其代谢物的低皮摩尔(Pm)浓度的方法。这项建议的第一个目标是开发一种预浓缩和除盐的方法，以允许检测低到亚皮摩尔水平的SP及其代谢物。这将通过评估几种不同类型的固相树脂和膜在从生物基质中保留低浓度SP的有效性来实现。本研究的第二个目标是将这种预浓缩/脱盐方法与毛细管电泳柱前衍生化或柱后衍生化的激光诱导荧光(LIF)检测相结合。一旦实现了这些目标，就有可能利用脑的微透析采样来研究SP通过血脑屏障的运输和代谢。以后的工作将比较开发的CE/LIF方法与毛细管液相色谱(CLC)和质谱仪(MS)检测所获得的检测限。未来的研究将致力于构建包含预浓缩/除盐装置、分离毛细管和检测系统的包容式微加工系统。

The ultimate goal of this research proposal is to develop a methodology for the measurement of low picomolar (pM) concentrations of substance P (SP) and its metabolites in highly ionic biological fluids. The first goal of this proposal is to develop a preconcentration and desalting method to permit the detection of low-to-subpicomolar levels of SP and its metabolites. This will be accomplished by evaluating several different types of solid phase resins and membranes for their effectiveness in retaining low concentrations of SP from biological matrixes. The second goal of this research is to use this preconcentration/desalting methodology in conjugation with capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection operating with either pre or postcolumn derivatization. Once these goals have been met, it will be possible to investigate the transport and metabolism of SP across the Blood Brain Barrier using microdialysis sampling of the brain. Later work will compare the detection limits achievable by the developed CE/LIF methodology to those obtainable by capillary liquid chromatography (CLC) with mass spectrometric (MS) detection. Future studies will work toward the construction of an inclusive microfabricated system that contains the preconcentration/desalting device, separation capillary, and detection system.